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- Table of Contents
DLL3 / Delta-like protein 3 · Western blot design guide
Design a Western Blot for DLL3
Real validated DLL3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DLL3 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
DLL3 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~64.6 kDa | |
| Observed band | ~65 kDa | |
| Gel | 5–20% (catalog PA1912) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DLL3 Western Blot Protocols
The PA1912 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, mouse brain, human HEK293 (catalog PA1912) |
| Gel % | 5–20% (catalog PA1912) |
| Load | 50ug; reducing conditions (catalog PA1912) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog PA1912) |
| Membrane | Nitrocellulose membrane (catalog PA1912) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog PA1912) |
| Primary antibody | PA1912 · 0.5 μg/mL (catalog PA1912) |
| Primary incubation | overnight at 4°C (catalog PA1912) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog PA1912) |
| Secondary incubation | 1.5 hour at RT (catalog PA1912) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1912) |
| Detection | ECL (catalog PA1912) |
Section 2
What Is the Expected DLL3 Western Blot Band Size?
DLL3 is predicted at 64.6 kDa and observed near 65 kDa; the cause of the small difference is not established.
| Band near 65 kDa | Matches the observed DLL3 band and is close to the 64.6 kDa predicted mass |
| Band slightly below 65 kDa | Could reflect removal of the signal peptide at residues 1–26; its migration effect is unmeasured |
| More than one band | Could reflect isoforms 1 and 2, although distinct band sizes are unconfirmed |
| Weak band in a soluble lysate fraction | DLL3 is a single-pass membrane protein and may be underrepresented in that fraction |
💡Expected DLL3 appearanceDLL3 has a predicted mass of 64.6 kDa and an observed band near 65 kDa; confirm band identity with an independent antibody or another appropriate control.
| UniProt predicted mass | 64.6 kDa, close to the observed band near 65 kDa |
| Signal peptide at residues 1–26 | Removal could make mature DLL3 smaller than its precursor; the band size after cleavage is not supplied |
| Isoform 1 | Its individual mass and migration relative to isoform 2 are not supplied |
| Isoform 2 | Its individual mass and migration relative to isoform 1 are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated DLL3 may be poorly recovered | Check membrane-protein extraction and test a membrane-enriched fraction |
| Band higher than expected | The identity or migration of the higher band is unestablished | Check reduction and verify DLL3 with an independent antibody |
| Band lower than expected | Signal-peptide removal could reduce mass, but the resulting band size is unknown | Verify the band with an independent antibody |
| Multiple bands | Isoforms 1 and 2 are listed, but distinct migration is unconfirmed | Verify band identity with an independent antibody and isoform-specific controls if available |
| Weak or no signal | Membrane-associated DLL3 may be underrepresented in the tested fraction | Check extraction and loading with a membrane-enriched fraction |
Experiment evidence
Western blot images showing DLL3 band
PA1912 · DLL3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DLL3 Western blot
🧪For positive controls for DLL3 in Western blot, you can use a validated DLL3-positive sample once identified, since the supplied HPA record provides no positive tissue or cell.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: The supplied HPA record identifies no tissue controls, so selecting a positive sample requires additional evidence.
HPA tissue expression evidence for DLL3
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced DLL3 Western Blot Tips
Deeper troubleshooting and optimisation questions for DLL3, answered from its protein features.
How should DLL3 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could DLL3 isoforms produce different bands?
Isoforms · DLL3 has two annotated isoforms. In isoform 2, canonical residues 587..618 are replaced by A. An antibody targeting that canonical segment may miss isoform 2. Check the antibody epitope before assigning a band to either isoform.
Do DLL3 features support a modification-related band shift?
PTM · No glycosylation site or modified residue is listed, although Ubl conjugation appears as a keyword. These annotations do not establish a visible shift or identify a modification site. Verify an additional band before attributing it to modification.
Does this guide establish induction of DLL3?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for DLL3 Western blot?
Transfer · DLL3 is a single-pass membrane protein with a predicted mass of 64.6 kDa. Check transfer near 65 kDa by examining both the membrane and residual gel. The supplied features do not specify an optimal membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA1912 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should DLL3 bands be quantified?
Quantitation · Use the same sample preparation and antibody epitope across samples, and quantify signals within the assay’s linear range. State whether the measurement covers one band or both annotated isoforms; their different C termini can affect detection by an epitope-specific antibody.
Why does DLL3 appear near 65 kDa?
Interpretation · The supplied apparent band near 65 kDa closely matches DLL3’s predicted 64.6 kDa. UniProt annotates a signal peptide at residues 1..26, but that feature alone does not establish a visible shift.
UniProt lists 18 disulfide bonds. Compare matched reducing and nonreducing samples if migration differs between preparations. A mobility change would not identify which bond or region caused it.
Check whether the antibody recognizes canonical residues 587..618, which differ in isoform 2. Compare matched reducing conditions given DLL3’s 18 annotated disulfide bonds, and confirm band identity with an independent epitope or targeted perturbation. The annotations alone cannot identify an unexpected band.
UniProt lists 18 disulfide bonds. Compare matched reducing and nonreducing samples if migration differs between preparations. A mobility change would not identify which bond or region caused it.
Check whether the antibody recognizes canonical residues 587..618, which differ in isoform 2. Compare matched reducing conditions given DLL3’s 18 annotated disulfide bonds, and confirm band identity with an independent epitope or targeted perturbation. The annotations alone cannot identify an unexpected band.
Boster reagents
DLL3 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Delta-like protein 3 DLL3 Antibody Picoband®
Cat # PA1912
The catalog reports PA1912, a rabbit polyclonal anti-DLL3 antibody with stated human reactivity. Its WB image shows an approximately 65 kDa band in rat brain, mouse brain, and human HEK293 lysates. The supplied evidence does not establish specificity through independent validation.
Which to pick: PA1912 is the only listed option and has a WB image. Choose it for a human sample when the reported HEK293 result fits your design; rat and mouse brain appear in the image, but those examples do not establish general species reactivity.
Source: BosterBio DLL3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NYJ7.
- Human Protein Atlas. DLL3 tissue expression.
- PMC5859037 — target-verified WB comparison
- PMC8973010 — target-verified WB comparison
- PMC11891050 — target-verified WB comparison